Abstract
The pseudofilamentous red alga Chroodactylon ornatum was grown in f/2 culture medium with the addition of 10, 34, and 100 μM (nominal concentration) cupric sulfate. The bioassays were terminated at two selected end points (days 4 and 12). Growth inhibition, changes in pigment composition, and oxidative stress indicators such as phenolic compounds and lipid peroxidation (dosed as thiobarbituric reactive substances) were observed in cultures with 34 and 100-μM cupric sulfate. Quinacrine (Atebrin) and chlorotetracycline fluorochromes showed abundant vacuoles of acidic content, related with mucilage secretion. Structural analyses by methylation, desulfation-methylation, alkaline treatment, and NMR spectroscopy revealed that the mucilaginous sheath of C. ornatum contains a sulfated galactan with a backbone of alternating 3-linked β-d-galactopyranose and 4-linked α-l-galactopyranose moieties, i.e., an agaran. The absence of 3,6-anhydrogalactose and of its precursor unit (α-galactose 6-sulfate) were confirmed. The highly sulfated polysaccharide contained these ester groups on the C-2 and C-4 hydroxyl groups of the 3-linked unit and on C-3 of the 4-linked units. The large proportion of sulfate esterification in this polysaccharide can be related to the extracellular biosorption of copper divalent cation (2.5 ± 0.4 mg per gram of dry weight) and to copper tolerance in bioassays.
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References
Andersen RA (ed) (2005) Algal culturing techniques. Elsevier Academic Press, San Diego
Andrade LR, Farina M, Amado Filho GM (2004) Effects of copper on Enteromorpha flexuosa (Chlorophyta) in vitro. Ecotoxicol Environ Safety 58:117–125
Areco MM, dos Santos Afonso M (2010) Copper, zinc, cadmium and lead biosorption by Gymnogongrus torulosus. Thermodyn Kinet Stud Colloids Surf B: Biointerfaces 81:620–628
Basson PW (1979) Marine algae of the Arabian Gulf coast of Saudi Arabia (second half). Bot Mar 22:65–82
Bermejo Román R, Alvárez-Pez JM, Acién Fernández FG, Molina Grima E (2002) Recovery of pure B-phycoerythrin from the microalga Porphyridium cruentum. J Biotechnol 93:73–85
Blaby Haas CE, Merchant SS (2012) The ins and outs of algal metal transport. Biochim Biophys Acta 1823:1531–1552
Bouzon ZL, Ouriques LC, Oliveira EC (2006) Spore adhesion and cell wall formation in Gelidium floridanum (Rhodophyta, Gelidiales). J Appl Phycol 18:287–294
Brown M, Newman JE (2003) Physiological response of Gracilariopsis longissima (Rhodophyceae) to sublethal copper concentrations. Aquat Toxicol 64:201–213
Byankina Barabanova AO, Sokolova EV, Anastyuk SD, Isakov VV, Isakov VV, Glazunov VP, Volod’ko AV, Yakovleva IM, Solov’eva TF, Yermak IM (2013) Polysaccharide structure of tetrasporic red seaweed Tichocarpus crinitus. Carbohydr Polym 98:26–35
Cases MR, Cerezo AS, Stortz CA (1995) Separation and quantitation of enantiomeric galactoses as their mono-O-methylethers as their diastereomeric 1-deoxy-1-(2′-hydroxypropylamino)-alditols. Carbohydr Res 269:333–341
Castilla JC (1996) Copper mine tailing disposal in Northern Chile rocky shores: Enteromorpha compressa (Chlorophyta) as a sentinel species. Environ Monit Assess 40:171–184
Ciucanu I, Kerek F (1984) A simple and rapid method for the permethylation of carbohydrates. Carbohydr Res 131:209–217
Contreras L, Moenne A, Correa JA (2005) Antioxidant responses in Scytosiphon lomentaria (Phaeophyceae) inhabiting copper enriched coastal environments. J Phycol 41:1184–1195
Correa JA, Castilla JC, Ramirez MA, Varas M, Lagos N, Vergara S, Moenne A, Roman D, Brown MT (1999) Copper mine tailings and their effects on marine algae in Northern Chile. J Appl Phycol 11:57–67
Diannelidis BE, Delivopoulos SG (1997) The effects of zinc, copper and cadmium on the fine structure of Ceramium ciliatum. Mar Environ Res 44:127–134
Dodgson KS, Price RG (1962) A note on the determination of the ester sulphate content of sulphated polysaccharides. Biochem J 84:106–110
Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for determination of sugars and related substances. Analyt Chem 28:350–356
Eklund BT, Kautsky L (2003) Review on toxicity testing with marine macroalgae and the need for method standardization––exemplified with copper and phenol. Mar Pollut Bull 46:171–181
Farías WR, Valente A, Pereira MS, Mourao PA (2000) Structure and anticoagulant activity of sulfated galactans. J Biol Chem 275:29299–29307
Gantt E (1990) Pigmentation and photoacclimation. In: Cole KM, Sheath RG (eds) Biology of the Red Algae. Cambridge University Press, Cambridge, pp 203–219
Genicot-Joncour S, Poinas A, Richard O, Potin P, Rudolph B, Kloareg B, Helbert W (2009) The cyclization of the 3,6-anhydro-galactose ring of iota-carrageenan is catalyzed by two d-galactose-2,6-sulfurylases in the red alga Chondrus crispus. Plant Physiol 151:1609–1616
Gloaguen V, Ruiz G, Morvan H, Mouradi-Givernaud A, Maes E, Krausz P, Strecker G (2004) The extracellular polysaccharide of Porphyridium sp.: an NMR study of lithium-resistant oligosaccharidic fragments. Carbohydr Res 339:97–103
Hader D, Lebert M, Sinha RP, Barbieri ES, Helbling EW (2002) Role of protective and repair mechanisms in the inhibition of photosynthesis in marine macroalgae. Photochem Photobiol Sci 1:809–814
Karsten U, West JA, Zuccarello GC, Nixdorf O, Barrow KD, King RJ (1999) Low molecular weight carbohydrate patterns in the Bangiophyceae (Rhodophyta). J Phycol 35:967–976
Karsten U, West JA, Zuccarello GC, Engbrodt R, Yokoyama A, Hara Y, Brodie J (2003) Low molecular weight carbohydrates of the Bangiophycidae (Rhodophyta). J Phycol 39:584–589
Kloareg B, Quatrano RS (1988) Structure of the cell walls of marine algae and ecophysiological functions of the matrix polysaccharides. Oceanogr Mar Biol Ann Rev 26:259–315
Kolender AA, Matulewicz MC (2002) Sulfated polysacchride from the red seaweed Georgiella confluens. Carbohydr Res 337:57–68
Küpper H, Etlík I, Spiller M, Küpper FC, Práil O (2002) Heavy metal-induced inhibition of photosynthesis: targets of in vivo heavy metal chlorophyll formation. J Phycol 38:429–441
Lapidot M, Shrestha RP, Weinstein Y, Arad Malis S (2010) Red microalgae: from basic know-how to biotechnology. In: Seckbach J, Chapman D (eds) Red algae in the genomic age. Springer, Netherlands, pp 205–225
Leonardi PI, Vasquez JA (1999) Effects of copper pollution on the ultrastructure of Lessonia spp. Hydrobiologia 398(399):375–383
Lewin RA, Robertson JA (1971) Influence of salinity on the form of Asterocytis in pure culture. J Phycol 7:236–238
Lichtenthaler H (1987) Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. Methods Enzymol 148:350–382
Lombardi AT, Vieira AH, Sartori LA (2002) Mucilaginous capsule adsorption and intracellular uptake of copper by Kirchneriella aperta (Chlorococcales). J Phycol 38:332–337
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Medcalf DG, Brannon JH, Scott JR, Allen GG, Lewis J, Norris RE (1981) Polysaccharides from microscopic red algae and diatoms. Proc Int Seaweed Symp 8:582–588
Mehta SK, Gaur JP (2005) Use of algae for removing heavy metal ions from wastewater: progress and prospects. Crit Rev Biotechnol 25:113–152
Miller IJ (1999) Further evaluation of the structure of the polysaccharide from Plocamium costatum with the use of set theory. Hydrobiologia 398/399:385–389
Müller KM, Lynch MD, Sheath RG (2010) Bangiophytes: from one Class to six; where do we go from here? In: Seckbach J, Chapman D (eds) Red algae in the genomic age. Springer, Netherlands, pp 241–259
Navarro DA, Stortz CA (2003) Determination of the configuration of 3,6-anhydrogalactose and cyclizable α-galactose 6-sulfate units in red seaweed galactans. Carbohydr Res 338:211–218
Navarro DA, Flores ML, Stortz CA (2007) Microwave-assisted desulfation of sulfated polysaccharides. Carbohydr Polym 69:742–747
Park JT, Johnson MJ (1949) A submicrodetermination of glucose. J Biol Chem 181:149–151
Pujals C (1961) Algunas observaciones sobre Asterocytis ornata (C. Ag.) Hamel, Rodofícea nueva para Argentina. Darwiniana 12:365–377 (in Spanish)
Ratkevicius N, Correa JA, Moenne A (2003) Copper accumulation, synthesis of ascorbate and activation of ascorbate peroxidase in Enteromorpha compressa (L.) Grev. (Chlorophyta) from heavy metal-enriched environments in northern Chile. Plant Cell Environ 26:1599–1608
Romay C, Gonzalez R, Ledon N, Remirez D, Rimbau V (2003) C-phycocyanin: a biliprotein with antioxidant, anti-inflammatory and neuroprotective effects. Curr Protein Pept Sci 4:207–216
Romera E, González F, Ballester A, Blázquez ML, Muñoz JA (2007) Comparative study of biosorption of heavy metals using different types of algae. Bioresour Technol 98:3344–3353
Santos JA, Mulloy B, Mourao PA (1992) Structural diversity among sulfated α-l-galactans from ascidians (tunicates). Eur J Biochem 204:669–677
Singleton VL, Orthofer R, Lamuela-Raventós RM (1999) Analysis of total phenols and other oxidation substrates and antioxidants by means of Folin-Ciocalteu reagent. In: Lester P (ed) Methods in Enzymology. Academic, London, pp 152–178
Statistica 5.5 (1999) Statsoft Inc, Tulsa, Oklahoma, USA
Stevenson TT, Furneaux RH (1991) Chemical methods for the analysis of sulphated galactans from red algae. Carbohydr Res 210:277–298
Stortz CA, Cerezo AS (2000) Novel findings in carrageenans, agaroids and ‘hybrid’ red seaweed galactans. Curr Top Phytochem 4:121–134
Sztrum AA, Sabatini SE, Rodríguez MC (2012) Isocitrate lyase activity and antioxidant responses in copper stressed cultures of Chlamydomonas reinhardtii (Volvocales, Chlorophyceae). Phycologia 51:135–143
Usov A (2011) Polysaccharides of red algae. Adv Carbohydr Chem Biochem 65:115–217
Vavilin D, Ducruet JM, Matorin DN, Venediktov PS, Rubin AB (1998) Membrane lipid peroxidation, cell viability and Photosystem II activity in the green alga Chlorella pyrenoidosa subjected to various stress conditions. J Photochem Photobiol B 42:233–239
Volesky B, Holan ZR (1995) Biosorption of heavy metals. Biotechnol Prog 11:235–250
Xia JR, Li YJ, Lu J, Chen B (2004) Effects of copper and cadmium on growth, photosynthesis, and pigment content in Gracilaria lemaneiformis. Bull Environ Contam Toxicol 73:979–986
Yoon HS, Müller KM, Sheath RG, Ott FD, Bhattacharya D (2006) Defining the major lineages of red algae (Rhodophyta). J Phycol 42:482–492
Yoon HS, Zuccarello GC, Bhattacharya D (2010) Evolutionary history and taxonomy of red algae. In: Seckbach J, Chapman D (eds) Red algae in the genomic age. Springer, Netherlands, pp 25–42
Zuccarello GC, West JA, Kikuchi N (2008) Phylogenetic relationships within the Stylonematales (Stylonematophyceae, Rhodophyta): Biogeographic patterns do no apply to Stylonema alsidii. J Phycol 44:384–393
Acknowledgments
This work was funded by grants from UBA (W-759) and CONICET (PIP 0559/10). JMC is a Research Fellow of the National Research Council of Argentina (CONICET), and CAS is a Research Member of the same institution.
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Cabrera, J.M., Stortz, C.A. & Rodríguez, M.C. A sulfated galactan from the mucilaginous sheath of the red filamentous alga Chroodactylon ornatum (Stylonematophyceae, Rhodophyta). J Appl Phycol 26, 1801–1811 (2014). https://doi.org/10.1007/s10811-014-0236-3
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DOI: https://doi.org/10.1007/s10811-014-0236-3